亚洲传统医药
亚洲传统医药

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›› 2015, Vol. 10 ›› Issue (3): 57-63.

• 论文 •    下一篇

A novel method of probe-melting curve for nucleic aciddetection without occupying fluorescence channel

Yang Han 1, Shangzhi Ji 2, Xiangzhong Ye 2, Yimin Li 2, Yixuan Zhang 1*   

  1. 1 The College of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University,Shenyang 110016, China;2 Department of research and development, Beijing Wantai Biological Pharmacy EnterpriseCo., Ltd, Beijing 102206, China
  • 收稿日期:2014-03-05 修回日期:2015-07-12 出版日期:2015-06-30 发布日期:2015-06-30
  • 通讯作者: Yixuan Zhang

A novel method of probe-melting curve for nucleic aciddetection without occupying fluorescence channel

Yang Han 1, Shangzhi Ji 2, Xiangzhong Ye 2, Yimin Li 2, Yixuan Zhang 1*   

  1. 1 The College of Life Science and Biopharmaceutics, Shenyang Pharmaceutical University,Shenyang 110016, China;2 Department of research and development, Beijing Wantai Biological Pharmacy EnterpriseCo., Ltd, Beijing 102206, China
  • Received:2014-03-05 Revised:2015-07-12 Online:2015-06-30 Published:2015-06-30
  • Contact: Yixuan Zhang

摘要： A novel method, probe-melting curve method, has been established to increase the detection throughputs within one fluorescence
channel. Besides using an asymmetric PCR assay, this method employs another specially designed probe with a 5′ terminus
modification. The method has been demonstrated by detection of rotavirus as model system. The sensitivity of probe-melting
curve method was proved to be equal to the conventional TaqMan real-time RT-PCR. And the experiment results were cross
checked by agarose gel electrophoresis. The successful establishment of probe-melting curve method breaks numbers of
fluorescence channels from the restrictions and enlarges the application fields of current real-time PCR instruments.

Abstract: A novel method, probe-melting curve method, has been established to increase the detection throughputs within one fluorescence
channel. Besides using an asymmetric PCR assay, this method employs another specially designed probe with a 5′ terminus
modification. The method has been demonstrated by detection of rotavirus as model system. The sensitivity of probe-melting
curve method was proved to be equal to the conventional TaqMan real-time RT-PCR. And the experiment results were cross
checked by agarose gel electrophoresis. The successful establishment of probe-melting curve method breaks numbers of
fluorescence channels from the restrictions and enlarges the application fields of current real-time PCR instruments.

Key words: Real-time PCR, RT-PCR, probe-based method, melting curve method

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